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目的：探讨活性氧过氧化氢（Ｈ２Ｏ２）对心肌细胞损伤作用。方法：用不同浓度的 Ｈ２Ｏ２作用于新生鼠培养心肌细胞，通过琼脂糖凝胶电泳、Ｇｉｅｍｓａ染色和流式细胞仪等方法观察Ｈ２Ｏ２对心肌细胞的凋亡性损伤作用，同时检测培养介质中乳酸脱氢酶（ＬＤＨ）和丙二醛（ＭＤＡ）等生化指标的改变。结果：５ｍｍｏｌ／Ｌ Ｈ２Ｏ２作用于培养心肌细胞６ｈ，在琼脂糖凝胶电泳上出现梯状ＤＮＡ带； Ｇｉｅｍｓａ染色细胞涂片可见到染色体浓聚或边缘化，成新月型聚集在核膜周边，并有凋亡小体的存在；流式细胞仪可检测出亚二倍体峰的出现。结论：Ｈ２Ｏ２具有浓度和时间依赖关系诱导心肌细胞的调亡，在低浓度Ｈ２Ｏ２（＜１ｍｍｏｌ／Ｌ）导致培养心肌细胞的早期生物化学改变，自由基含量升高，膜通透性增加，同时伴有少量心肌细胞调亡；而在高浓度Ｈ２Ｏ２（＞１０ｍｍｏｌ／Ｌ）时会造成培养心肌细胞的坏死性死亡，细胞膜崩塌，脂质过氧化物（ＭＤＡ）大量产生，ＬＤＨ大量泄漏，ＤＮＡ琼脂糖凝胶电泳出现弥散（ｓｍｅａｒ）泳带；５～１０ ｍｍｏｌ／Ｌ Ｈ２Ｏ２诱导大量心肌细胞凋亡性死亡，同时伴有 ＬＤＨ和 ＭＤＡ含量升高等生物化学的改变。
Objective: To investigate the effect of reactive oxygen species hydrogen peroxide (H2O2) on cardiomyocyte injury. Methods: Cardiomyocytes were cultured with different concentrations of H2O2 in neonatal rats. The injury of H2O2 on cardiomyocytes was observed by agarose gel electrophoresis, Giemsa staining and flow cytometry. Meanwhile, Hydrogenase (LDH) and malondialdehyde (MDA) and other biochemical indicators of change. Results: The cardiomyocytes were cultured with 5mmol / L H2O2 for 6h, and ladder DNA bands appeared on agarose gel electrophoresis. Giemsa stained cells showed clustering or marginalization of chromosomes, And the existence of apoptotic bodies; flow cytometry can detect the appearance of sub-diploid peaks. CONCLUSION: H 2 O 2 induces cardiomyocyte apoptosis in a concentration-dependent manner in time-dependent manner. H2O2 (<1 mmol / L) resulted in early biochemical changes of cardiomyocytes, increased free radical content and increased membrane permeability A small amount of cardiomyocytes apoptosis; while in high concentration of H2O2 (> 10mmol / L) will cause necrosis of cultured myocardial cells, cell membrane collapse, a large number of lipid peroxidation (MDA), LDH leakage, DNA agarose The gel electrophoresis showed a smear band. 5 ~ 10 mmol / L H 2 O 2 induced a large number of apoptotic cardiomyocytes with biochemical changes such as elevated LDH and MDA contents.